The influence of Mg<sup>2+</sup>/adenine nucleotide ratios and absolute concentration of Mg<sup>2+</sup>/adenine nucleotide on the observed velocity of some kinase reactions

Garner, Pamela S.; Rosett, Theodore

doi:10.1016/0014-5793(73)80803-8

FEBS Letters

1973

DOI: 10.1016/0014-5793(73)80803-8

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The influence of Mg²⁺/adenine nucleotide ratios and absolute concentration of Mg²⁺/adenine nucleotide on the observed velocity of some kinase reactions

Pamela S. Garner

Theodore Rosett

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Cited by 19 publications

(9 citation statements)

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“…3). This (26,27). This suggests that the reaction that regulates the onset of DNA synthesis has a similar Mg2+ dependency, and that it might be possible to identify it by this property.…”

mentioning

confidence: 86%

Magnesium reverses inhibitory effects of calcium deprivation on coordinate response of 3T3 cells to serum

Rubin

Terasaki

Sanui

1978

Proc. Natl. Acad. Sci. U.S.A.

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When animal cells are stimulated to multiply by serum, hormones, or various other unrelated agents added to their culture medium, several early events occur, including acceleration in transport and phosphorylation of substrates (1, 2), energy metabolism (3), and synthesis of differentiated cell products (4, 5). After an extended lag period, the accelerated onset of DNA synthesis becomes manifest (6). This array of events has been designated the coordinate response to distinguish it from the "pleiotypic response," which explicitly excludes differentiated functions (7). We have shown that all the elements of the coordinate response can be inhibited in chick embryo cells in a reversible manner by lowering the [Mg2+] of the medium (8,9). From these results and several other considerations, we have proposed that variations in the availability of Mg2+ for transphosphorylation reactions within the cell mediate the coordinate response of cells to external effectors (8). Others have proposed that Ca2+ plays a central role in mediating the growth response of cells to external effectors (10). Support for a mediating role for Ca2+ is chiefly based on the observation that drastically lowering the [Ca2+] of the medium inhibits the onset of DNA synthesis and thereby limits the multiplication of cells (11). Additional support seemed to come from the finding that supranormal concentrations of Ca2+ stimulate the growth of quiescent 3T3 cells (12), but this has proven to be a nonspecific effect caused by insoluble complexes of Ca2+ and inorganic orthophosphate (Pi) (13,14). It has also been shown that Ca2+ deprivation in chick embryo cells causes a marked increase in the passive permeability of cells (15) The procedures for measuring cation contents of cells by atomic absorption spectrophotometry were previously reported (20). Culture dishes (100 mm) were used in the cation determination experiments. Briefly, the cultures were washed five times with 10 ml per wash of C02-free 0.25 M sucrose solution, 4379The publication costs of this article were defrayed in part by page charge payment. This article must therefore be hereby marked "advertisement" in accordance with 18 U. S. C. §1734 solely to indicate this fact.

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“…3). This (26,27). This suggests that the reaction that regulates the onset of DNA synthesis has a similar Mg2+ dependency, and that it might be possible to identify it by this property.…”

mentioning

confidence: 86%

Magnesium reverses inhibitory effects of calcium deprivation on coordinate response of 3T3 cells to serum

Rubin

Terasaki

Sanui

1978

Proc. Natl. Acad. Sci. U.S.A.

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“…The (19). The decrease in protons that occurs with increased pH should shift HATP3-, which is inhibitory for many phosphorylation reactions, to MgATP2-, which is the substrate for these reactions (20,21). Thus, protein synthesis could be affected both directly and indirectly by altered pH through its effect on the distribution of magnesium among adenine nucleotides.…”

mentioning

confidence: 99%

Evidence that intracellular magnesium is present in cells at a regulatory concentration for protein synthesis.

Terasaki¹,

Rubin²

1985

Proc. Natl. Acad. Sci. U.S.A.

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When extracellular magnesium is reduced by a factor of 50 (from 1.0 to 0.02 mM), the total intracellular magnesium of a spontaneously transformed clone of 3T3 cells decreases by 30-50%. Protein synthesis rates in these cells were measured as the intracellular magnesium decreased. Protein synthesis rates and magnesium content were found to decrease in parallel with each other. At 3 hr, a decrease to 84% ofcontrol values of magnesium content was accompanied by a decrease to 85% of control values of leucine incorporation rates. A larger inhibition had occurred by 12 hr, when the magnesium had decreased to 67% and leucine incorporation rates had decreased to 57%. When magnesium was restored to maiumdeprived cells, both magnesinm content and leucine incorporation increased about 2-fold by 1 hr. In the experiments reported here, initial small changes in magnesium content are associated with changes in protein synthesis rates. This strongly suggests that magnesium is present at a regulatory rather than excess concentration for protein synthesis. The results are consistent with a role for intracellular magnedum in the regulation of protein synthesis and support the hypothesis that magnesium has a central role in the regulation of metabolism and growth.Protein synthesis rates in in vitro systems are known to be affected by magnesium (1). However, there has been little work devoted to characterizing the effects of magnesium on protein synthesis in intact cells. Perhaps the main reason is that magnesium is difficult to vary within cells-increases or decreases by a factor of 10 of extracellular magnesium have no effect on the total intracellular magnesium contents of the cell (2).In an earlier paper, intracellular magnesium of 3T3 cells was varied by lowering the extracellular calcium concentration to 0.02 mM and then varying extracellular magnesium from 1 to 20 mM and to even higher inhibitory magnesium concentrations (3 ity, 51.6 Ci/mmol; 1 Ci = 37 GBq) was used at an activity of 2 ,uCi/ml in 2 ml of medium for the 60-mm dishes.[3H]Thymidine (original specific activity, 20 Ci/mmol) was used at an activity of 1 puCi/ml in 2 ml of medium. The length of the labeling period is given in the figure legends. All labeling media were made using medium of the same composition as the growth medium of the cells at the time of the label. At the end of the labeling period, cultures were washed three times with ice-cold Tris-buffered saline, extracted with ice-cold 5% trichloroacetic acid for 10 min, washed twice with trichloroacetic acid, and then solubilized in 2 ml of 0.2 M NaOH. Samples of the NaOH were taken for scintillation counting and protein determination. Duplicate protein determinations were run.Ion Measurements. The method of Sanui and Rubin (8) was used. Briefly, cultures were first washed three times with C02-free 0.25 M sucrose (pH 7). This was followed by one wash with carbonated 0.25 M sucrose (pH 4), whose purpose was to displace externally bound cations by protons. The cells were scraped into distilled deioni...

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“…Morrison and Heyde have pointed out that this is a function of type of buffer, ionic strength, pH, and the presence of other enzymes [4]. This question becomes even more important since the availability of Mg2+ in vivo appears to be an important control [5,6].…”

mentioning

confidence: 99%

Computer Simulation Study of Hexokinase II from Ehrlich Ascites Cells

Garfinkel

1975

Eur J Biochem

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A study of the mechanism of hexokinase II from ascites cells and the effects of its binding to mitochondrial membranes has been carried out by computer simulation. This is based on experimental data of Kosow and Rose and of Gumaa and McLean, and the theoretical methods of Cleland. For the soluble enzyme the mechanism is random with ternary product‐inhibition complexes; when bound to mitochondria, the mechanism becomes ordered‐on, random‐off, as the binding of ATP to the free enzymes becomes negligibly slow. The requirements of experimental data for mechanistic studies are discussed.

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The influence of Mg²⁺/adenine nucleotide ratios and absolute concentration of Mg²⁺/adenine nucleotide on the observed velocity of some kinase reactions

Cited by 19 publications

References 15 publications

Magnesium reverses inhibitory effects of calcium deprivation on coordinate response of 3T3 cells to serum

Magnesium reverses inhibitory effects of calcium deprivation on coordinate response of 3T3 cells to serum

Evidence that intracellular magnesium is present in cells at a regulatory concentration for protein synthesis.

Computer Simulation Study of Hexokinase II from Ehrlich Ascites Cells

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The influence of Mg2+/adenine nucleotide ratios and absolute concentration of Mg2+/adenine nucleotide on the observed velocity of some kinase reactions

Cited by 19 publications

References 15 publications

Magnesium reverses inhibitory effects of calcium deprivation on coordinate response of 3T3 cells to serum

Magnesium reverses inhibitory effects of calcium deprivation on coordinate response of 3T3 cells to serum

Evidence that intracellular magnesium is present in cells at a regulatory concentration for protein synthesis.

Computer Simulation Study of Hexokinase II from Ehrlich Ascites Cells

Contact Info

Product

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The influence of Mg²⁺/adenine nucleotide ratios and absolute concentration of Mg²⁺/adenine nucleotide on the observed velocity of some kinase reactions